Methods for concentration and extraction of lubricity compounds and biologically active fractions from naturally derived fats, oils and greases

ABSTRACT

Methods for recovery of concentrates of lubricating compounds and biologically active compounds from vegetable and animal oils, fats and greases that allow separation of triglycerides, from components with higher lubricity or biological activity or enrichment protocols that increase the concentration of high lubricity or biologically active compounds in the triglyceride. The triglycerides are transesterified with a lower alcohol to produce alkyl esters. Following the conversion process the esters are separated from high molecular weight high lubricity compounds and biologically active compounds by distillation. The esters have some lubricity and may be sold as pollution reducing fuel components. The high boiling point compounds that are the residues of distillation, however, can either contribute significant lubricity and may be used widely in lubricant applications or added to petroleum fuels to decrease friction or the biologically active components may be used in nutritional, cosmetic and therapeutic applications. Therapeutic applications include use in human diets to lower cholesterol.

CROSS REFERENCE TO RELATED APPLICATION

This application is a Continuation-in-part of previously filed U.S.patent application Ser. No. 11/290,781 filed 1 Dec. 2005

FIELD OF INVENTION

The present invention relates to methods for producing a high lubricityfraction and for producing bioactive fractions from fats, oils andgreases derived from a wide variety of animal and vegetable sources. Inthis specification, the terms “oils, fats and greases” are usedsynonymously to describe starting materials derived from vegetable andanimal sources. Oils tend to be liquid at room temperature and arederived from many biological sources such as whales, fish and oil seed.Fats are generally solid at room temperature and are derived from thesame sources as oils. Greases usually have high melting points and theymay be synthetic products. Some synthetic greases are plant derived,others are from animals. The novel methods either separate lowerlubricity components of the fat, oil, or grease from higher lubricityfractions or enrich the concentration of high lubricity components orcombine extraction and enrichment. In a preferred embodiment the lowerlubricity components are made volatile by chemical reactions that splitthe triglyceride component of fat, oil, or grease. These reactions mayproduce industrially useful products such as fatty acid methyl esters,fatty acids, fatty alcohols, fatty aldehydes or fatty amides of theoriginal fat, oil, or grease which may be separated from the higherlubricity components by distillation. The lower lubricity componentsfrom fat splitting have inherent value that is not diminished by theseparation of the high lubricity fraction. In fact, the low lubricityfraction may have increased value as a result of the separation. Thehigh lubricity fraction is a collection of higher molecular weightsubstances present in the fat, oil or grease or a modified componentthereof. In another preferred embodiment the high lubricity component ofthe fat, oil or grease is separated from the triglyceride by absorptiononto a solid phase medium. Depending on the nature of the solid phaseextraction medium either the lower lubricity components or the higherlubricity components are preferentially bound to the solid phaseextraction medium. The concentrate is then recovered from the solidphase by extraction or from the liquid phase by evaporation. In afurther preferred embodiment the separation of higher lubricity andlower lubricity components is achieved by crystallisation from asolvent. In another embodiment of the present invention the novelmethods separate triglyceride components of the fat, oil, or grease frombiologically active fractions. The methods also enrich the concentrationof biologically active components in a selective extraction process. Ina preferred embodiment the glyceride components are made volatile bychemical reactions that split the oil triglyceride. These reactions mayproduce industrially useful products such as fatty acids, fatty acidesters, fatty alcohols, fatty aldehydes or fatty amides of the originalvegetable oil which may be separated from the biologically activecomponents by distillation. The distilled components from fat splittinghave inherent value that is not diminished by the separation of thebiologically active fraction. In fact, the distilled components may haveincreased value as a result of the separation. The biologically activefraction is a collection of higher molecular weight substances presentin the starting material.

Extraction procedures may also be manipulated to improve the content ofcompounds that impart lubricity to the fat, oil or grease. In apreferred embodiment canola seed is mechanically pressed to remove oilthat has lower levels of the desired high lubricity compounds.Mechanical extraction of the seed is followed by solvent extraction thatproduces oil with a surprising level of lubricity. The lubricity isimparted through the high ratio of lubricity enhancing products totriglyceride extracted with the oil.

Extraction procedures may also be manipulated to improve the content ofbiologically active compounds. In a preferred embodiment canola seed ismechanically pressed to remove oil that has lower levels of the desiredbiologically active compounds. Mechanical extraction of the seed isfollowed by solvent extraction of the solids in a process that producesoil with a surprising level of biologically active components.

Surprisingly it has also been discovered that specific fractions ofoil-bearing material may be selected that possess higher levels ofbiologically active components. In a preferred embodiment small seed isselected prior to extraction to enable recovery of greater levels of thebiologically active component. The invention includes the selection ofthese materials by physical and other methods.

BACKGROUND OF THE INVENTION

Since 1993, environmental legislation in the U.S. has required that thesulfur content of diesel fuel be less than 0.05%. In 2007 the sulfurcontent of diesel has been legislated to contain less than 15 ppmsulfur. The reduction in the sulfur content of diesel fuel has resultedin lubricity problems. It has become generally accepted that thereduction in sulfur is also accompanied by a reduction in polaroxygenated compounds and polycyclic aromatics includingnitrogen-containing compounds responsible for the reduced boundarylubricating ability of severely refined (low sulfur) fuels. While lowsulfur content is not in itself a lubricity problem, it has become themeasure of the degree of refinement of the fuel and thus reflects thelevel of the removal of polar oxygenated compounds and polycyclicaromatics including nitrogen-containing compounds.

Low sulfur diesel fuels have been found to increase the sliding adhesivewear and fretting wear of pump components such as rollers, cam plate,coupling, lever joints and shaft drive journal bearings.

Concern for the environment has resulted in moves to significantlyreduce the noxious components in emissions when fuel oils are burnt,particularly in engines such as diesel engines. Attempts are being made,for example, to minimize sulfur dioxide emissions by minimizing thesulfur content of fuel oils. Although typical diesel fuel oils have inthe past contained 1% by weight or more of sulfur (expressed aselemental sulfur) it is now mandatory to reduce the sulfur content toless than 15 ppm (0.0015%).

Additional refining of fuel oils, necessary to achieve these low sulfurlevels, often results in a reduction in the levels of polar components.In addition, refinery processes can reduce the level of polynucleararomatic compounds present in such fuel oils.

Reducing the level of one or more of the sulfur, polynuclear aromatic orpolar components of diesel fuel oil can reduce the ability of the oil tolubricate the injection system of the engine. As a result of poor fuellubrication properties the fuel injection pump of the engine may failrelatively early in the life of an engine. Failure may occur in fuelinjection systems such as high-pressure rotary distributors, in-linepumps and injectors. The problem of poor lubricity in diesel fuel oilsis likely to be exacerbated by future engine developments, aimed atfurther reducing emissions, which will result in engines having moreexacting lubricity requirements than present engines. For example, theadvent of high-pressure unit injectors is anticipated to increase thefuel oil lubricity requirement.

Similarly, poor lubricity can lead to wear problems in other mechanicaldevices dependent for lubrication on the natural lubricity of fuel oil.

Lubricity additives for fuel oils have been described in the literature.WO 94/17160 describes an additive, which comprises an ester of acarboxylic acid and an alcohol, wherein the acid has from 2 to 50 carbonatoms and the alcohol has one or more carbon atoms. Glycerol monooleateis an example. Although general mixtures were contemplated, no specificmixtures of esters were disclosed.

U.S. Pat. No. 3,273,981 discloses a lubricity additive being a mixtureof A+B wherein A is a polybasic acid, or a polybasic acid ester made byreacting the acid with C₁-C₅ monohydric alcohols; while B is a partialester of a polyhydric alcohol and a fatty acid, for example glycerylmonooleate, sorbitan monooleate or pentaerythitol monooleate. Themixture finds application in jet fuels.

U.S. Pat. No. 6,080,212 teaches of the use of two esters with differentviscosity in diesel fuel to reduce smoke emissions and increase fuellubricity. In one preferred embodiment of that invention methyloctadecenoate, a major component of biodiesel, was included in theformula. Similarly, U.S. Pat. No. 5,882,364 also describes a fuelcomposition comprising middle distillate fuel oil and two additionallubricating components. Those components being (a) an ester of anunsaturated monocarboxylic acid and a polyhydric alcohol and (b) anester of a polyunsaturated monocarboxylic acid and a polyhydric alcoholhaving at least three hydroxy groups.

The approach of using a two component lubricity additive was pioneeredin U.S. Pat. No. 4,920,691. The inventors describe an additive and aliquid hydrocarbon fuel composition consisting essentially of a fuel anda mixture of two straight chain carboxylic acid esters, one having a lowmolecular weight and the other having a higher molecular weight.

In U.S. Pat. No. 5,713,965 the synthesis of alkyl esters from animalfats, vegetable oils, rendered fats and restaurant grease is described.The resultant alkyl esters are reported to be useful as additives toautomotive fuels and lubricants.

Alkyl esters of fatty acids derived from vegetable oleaginous seeds wererecommended at rates between 100 to 10,000 ppm to enhance the lubricityof motor fuels in U.S. Pat. No. 5,599,358. Similarly a fuel compositionwas disclosed in U.S. Pat. No. 5,730,029 comprising low sulfur dieselfuel and esters from the transesterification of at least one animal fator vegetable oil triglyceride.

Most commercially available plant oils are highly enriched intriacylglycerol and diacyl glycerols. However, as well as includingthese more abundant substances, plant oils are known to contain a largenumber of biologically active components. While the biologically activecomponents may occur at concentrations sufficient to impart usefulbiological responses their concentrations are often insufficient formany applications.

Phytosterols are known by those skilled in the art as dietary materialsthat can lower blood serum cholesterol. In fact knowledge that dietaryphytosterols decrease cholesterol extend back to 1951 (Peterson, Procsoc Exp Biol Med 1951; 78:1143). Jones et al. (Can J Physiol Pharmacol1997; 75:217) reports that phytosterols are consumed at a level of200-400 mg/day. However clinical effects described in many publicationsare significant when phytosterols or their esters are utilised atconcentrations well above the natural concentrations found in vegetableoils. For example Shin et al. (Nutritional Research 2003; 23:489)provided human test subjects with a beverage containing 800 mg/servingand with 2-4 servings/day. The eight-week protocol significantly loweredcholesterol in the test population.

Sterols occur at significant concentrations in many vegetable oilsmainly as free sterols and as their fatty esters. Nevertheless, theconcentrations found in most sources are less than sufficient to producea therapeutic effect.

Meguro et al. (Nutrition 2003; 19:670-675) report that diacylglycerolsinteract with sterol provided in the diet to reduce cholesterol levelsin New Zealand White (NZW) rabbits below that achieved by the samecontent of sterol in triacyl glycerol. They hypothesise that the diacylglycerol interacts with the sterol partially through the highersolubility of the sterol in the diacyl glyceride phase.

Dolichol is a naturally occurring high molecular weight alpha-saturatedpolyprenol that is widely distributed in living organisms. Mammalssynthesise dolichol in normal metabolism but may take it up from thediet as well (Jacobsson et al. 1989; FEBS 255:32). U.S. Pat. No.4,599,328 teaches that dolichol is an effective treatment forhyperuricuria, hyperlipemia, diabetes and hepatic disease. It has alsobeen demonstrated in animal model systems that dolichol and dolicholphosphate can act as antihypertensive treatments (U.S. Pat. No.4,175,139).

Polyisoprenol compounds are similar to dolichol in structure but serve adifferent function in metabolism. Polyisoprenol compounds are widelydistributed and known to be components of many vegetable oils.

Tocols are an important class of nutrients and includes the essentialnutrient vitamin E or alpha tocopherol. While vitamin E has a wide rangeof metabolic functions that are realised at low rates of incorporationin the diet supplementation with vitamin E is believed to have potentialbenefits in the prevention of ageing and disease. While vegetable oilsare significant sources of vitamin E in the diet levels may beinadequate to meet recommended daily allowances and recommended levelsfor therapeutic effects.

Plant oils also contain chromanols including ubiquinone, ubiquinol,plastoquinone and plastoquinol. These compounds are potent antioxidantsand are thought to slow ageing processes.

Carotenoids and notably lutein and zeazanthin are important constituentsof certain vegetable oils. Consumption of these carotenoids has beenassociated with the prevention of specific eye diseases. For example, aninverse association has been noted with the incidence of advanced,neovascular, age-related macular degeneration (AMD) and the dietaryintake of lutein and zeaxanthin. Individuals whose diets are modified toinclude an increased intake of lutein and zeaxanthin generally respondwith an increase in concentrations in these pigments in their serum andmaculae (Hammond et al. 1997; Invest. Opthamol. Vis. Sci. 38:1795).

Typically phytosterol and vitamin E are obtained from industrial streamsencountered in the processing of plant based oils. A phytosterol andtocopherol rich fraction is recovered during the refining of vegetableoil where in a late stage of refining vegetable oil is steam distilledunder vacuum to deodorise the oil. The deodoriser concentrate is rich infree fatty acid, free sterol and tocopherol and substantially devoid ofsterol ester, dolichol, diacylglycerol and carotenoids. This fraction isa major source of sterol and tocopherol used in nutritionalapplications.

Phytosterol is also derived from the pulp and paper industry wheresolution from alkali washed wood pulp is acidified to produce a complexmixture of plant lipids known as tall oil. This latter fraction can bedivided to produce fatty acids, rosin acids and sterols.

Carotenoids used for dietary purposes may be derived from a number ofsources. For example, marigold may be harvested and processed as asource of dietary lutein. Other dietary carotenoids, includingastaxanthin and canthaxanthin are synthesised by classical organicsynthetic methods.

While vegetable oils may be rich sources of sterol esters, tocols, andcarotenoids methods of recovery of these components are inefficient andproducts must be fractionated and reformulated for use.

SUMMARY OF THE INVENTION

It is known by those skilled in the art that fuel additives that enhancelubricity may be produced that contain lower alkyl esters of fats, oilsand greases yet surprisingly it is revealed, in the present invention,that these mixtures contain ingredients with substantially higherlubricity. Furthermore methods are disclosed to efficiently recoverthese high lubricity components. In preferred methods the triglyceridecomponents of the fat, grease or oil are converted to lower molecularweight compounds such as fatty acids, fatty amides or fatty acid alkylesters. In forming the lower molecular weight compound it becomespossible to readily separate the bulk material from the high lubricitycomponents by distillation. In a preferred embodiment the fat, oil orgrease is transesterified to produce a lower alkyl ester using methodsknown to those skilled in the art. The ester is then distilled andrecovered for other purposes and the column bottoms of distillation arerecovered and refined to remove free acids formed in distillation. Therefined column bottoms recovered from the distillation have substantialefficacy as lubricity additives. In a preferred embodiment the fat, oilor grease is converted to fatty acids. The fatty acids are thendistilled and recovered for other purposes and the column bottoms ofdistillation are recovered and refined to remove residual free acidsformed in distillation. The refined column bottoms also have substantialefficacy as lubricity additives. The lubricity concentrate comprises acomplex mixture of phospholipid, sterol, tocol, quinone, polyisopreneand polyisoprenol and other lipid soluble components. In a preferredembodiment of the present invention where the concentrate is an enrichedconcentrate of lipid substances with molecular weights greater than 400.

While the present invention may be accomplished through fat splitting orother chemical modification followed by crystallisation or distillationas preferred methods of concentrating the lubricity fraction, othermethods of concentrating specific classes of oil soluble compounds fromtriglyceride are also acceptable. For example, those skilled in the artwill recognise that it is possible to recover enriched fractions fromfats, oils and greases by solid phase extraction and chromatographicmethods. Solid phase extraction may be combined with chemicalmodification steps or the chemical modification may be forgone in theprocess of preparing the high lubricity concentrates.

Furthermore we have made the surprising discovery that the method ofprocessing the oil may also act to concentrate the oil solublecomponents that impart lubricity. Processing conditions may be modifiedto enhance the extraction of high lubricity minor components of oilseedand animal fat. The present invention includes pre-extraction treatmentsthat enhance either or both the concentration of high lubricitycomponents in oils.

In another preferred embodiment of the present invention where theconcentrate is enriched in dolichol, other polyisoprenols and theirderivatives, and the present invention describes methods ofoptimally-preparing concentrates of biologically active oil solublecompounds. In the preferred art the triglyceride components of vegetableoils are subject to chemical rearrangements to form new products thathave a lower molecular weight and boiling point. Reaction conditions areselected so as to prevent the degeneration of the biologically activecomponents. It has been found that the process of distillation undermild conditions can remove much of the modified glyceride productleaving behind a concentrate of biologically active substances. As mostplant oils are sources of carotenoid, phytosterol, tocol, chromanol, anddolichol and these components have relatively high molecular masses itis common to find these compounds present in the concentrate.

In a preferred embodiment ethyl esters were synthesised using analkaline catalyst reaction of ethanol with low erucic acid rapeseed oil,a plant oil that is highly rich in triglyceride. In this embodiment thereaction conditions are maintained under the mildest possible conditionsto prevent the destruction of the biologically active components. Afterthe reaction the glycerol released in the reaction and excess ethanolwere removed, the esters were distilled in a thin film still to recoverover 90 percent of the ethyl ester as a concentrate. The resultingconcentrate was highly enriched in phytosterol, tocol, dolichol andcarotenoid.

The instant invention also includes methods of pre-extraction thatproduce enriched concentrates of biologically active compounds. In apreferred embodiment low erucic acid rapeseed was crushed mechanicallyusing a commercial expeller press under mild conditions to recover anoil fraction that had reduced levels of biologically active components.The mild conditions of mechanical extraction are known to those skilledin the art as cold pressing. After mechanical extraction the solidfraction was subject to solvent extraction to recover the remaining oil.The second oil possessed elevated concentrations of many biologicallyactive components including phytosterol, tocol, dolichol and carotenoid.Although the triglyceride remained a major component of the solventextracted oil the concentration step allowed for the use of moreefficient process steps in the production of a concentrate ofbiologically active components. It is a particular benefit of thislatter preferred embodiment that the manufacturing process generates asignificant fraction of oil that has not been extracted by utilising asolvent.

DETAILED DESCRIPTION OF THE INVENTION

Vegetable oils, such as tall, soybean, canola, palm, sunflower, hemp,rapeseed, flaxseed, corn or coconut, are a complex mixture of molecularcomponents of which triglycerides are usually the most abundantcomponent. Numerous other seed oils are known and are also included inthis invention. Palm and olive oil are derived by processing the fruitsof the palm and olive trees. Tall oil is a vegetable oil recovered fromthe pulp and paper industry and is essentially the oil present in wood.Similarly, animal fats and greases, such as those derived from swine,poultry and beef, are predominantly triglyceride in composition.Triglycerides are triesters of glycerol and carboxylic acids that havegreat industrial importance. In industry triglycerides are reacted withwater to form fatty acids, hydrogen to form fatty alcohols, reducingagents to form aldehydes, amines to form fatty amides and alcohols toform alkyl esters. Triglycerides have relatively high molecular weights,usually greater than 800 amu and thus are difficult to distill. However,fatty acids, fatty amides, fatty alcohols and fatty alkyl esters oflower alcohols have lower molecular weights and are readily distilledunder vacuum. The residue left after vacuum distillation is aconcentrate of substances with molecular weights above those of thefatty acid, amide, alcohol, aldehyde or ester.

Preconcentration

The oilseeds are typically processed both by mechanical and solventextraction to recover the seed oil. Mechanical extraction methodsinclude hydraulically operated oil presses, continuous screw presses,and extruders adapted for oil extraction. Mechanical extraction methodsmobilise a portion of the oil by both shear and pressure which rupturesoil containing structures in the seed. Once the oil is mobilised it mayflow away from the solids which are held in the press by physicalstructures such as metal bars. Depending on the severity of thepressure, temperature and shear conditions the amount of oil recoveredfrom oilseed varies. In order to maximise the yield of oil it ispossible to utilise more severe extraction conditions. It is common tothose skilled in the art to utilise expeller presses in sequence tofirst remove a portion of the oil under milder extraction conditionsthen to follow this by a second expeller press treatment under moresevere conditions. It is an example of the current art where the totalpressed oil is utilised for recovery of biologically active components.It is a preferred embodiment of the present invention that the oilrecovered from the second oilseed press is utilised as a superior sourcefor the biologically active materials. In advanced expeller pressdesigns it is common to increase the severity of pressing of the oilseedmaterial as it passes along the press. Oil recovered from the earlyportion of the press is extracted under milder conditions than materialrecovered from the latter stages of the press. Surprisingly it has beenfound that the level of biologically active oil soluble ingredients isenriched in the oil recovered in the latter stages of pressing. It is apreferred embodiment of the present invention that the oil recoveredfrom the latter stages of a press is recovered and utilised forextraction of the biologically active fraction. It is also commonpractise in industry to utilise an expeller press to remove a portion ofthe oil followed by placing the partially deoiled seed meal in acontinuous or batch solvent extraction vessel. The seed meal may then befully deoiled by extracting with a suitable non-polar solvent. Usefulsolvents include but are not limited to hexane, supercritical carbondioxide, propane, ethanol, isopropanol and acetone. It is an embodimentof the present invention that oil recovered by solvent extraction,following mechanical removal of the oil is utilised as a superior sourceof the biologically active materials.

Molecular Weight Reduction: Transesterification

Once the oil has been separated, it is an object of the currentinvention to produce a useful concentrate of the biologically activefraction. In order to concentrate the biologically active molecules itis necessary to separate them from the higher molecular weight and oftenless biologically active triglyceride materials as they may constituteover 95 percent of the seed oil. Typical seed oil glycerides havemolecular masses of greater than 800 g/mole. As such these compounds aredifficult to distill. In the current art to achieve this separation itis necessary to convert the triglyceride oils to lower molecular weightforms so that they are readily distilled to leave a residue of thebiologically active concentrate.

Glycerides are esters of glycerol and they are readily reacted toproduce fatty compounds that have lower molecular weight than the parentglyceride. In a preferred embodiment of the current invention theglyceride component of the seed oil is converted to fatty acid esters.There are many documented approaches to the chemical conversion oftriglycerides to alkyl esters known by those skilled in the art and suchapproaches other than those described herein are included in the instantinvention. In a preferred embodiment vegetable oil that containsbiologically active compounds is treated with a solution of an alkalibase, such as potassium hydroxide dissolved in ethanol under anhydrousconditions. The ensuing reaction converts the triglyceride to thecorresponding ethyl ester. After conversion, the molecular weight of thefatty ester compounds is substantially reduced while the biologicallyactive components with higher molecular weights are not similarlyreduced in molecular mass. Distillation will selectively remove thefatty ester compounds and leave a unique residue of biologically activematerials with higher molecular weights. While the use of distillationis preferred for separation of the alkyl ester component of the reactionit is obvious to one skilled in the art that other methods of separatingmolecules that differ in size that could be used to separate the alkylesters from the biologically active fraction. These methods are includedin the present invention. As the products of the current invention maybe produced using ethanol, the use of other lower alkanols with between1 and 5 carbon atoms is included as a portion of the current art.

Molecular Weight Reduction: Hydrolysis

In a preferred embodiment of the current invention the glyceridecomponent of the seed oil is converted to fatty acids. There are manydocumented approaches to the chemical conversion of triglycerides tofatty acids known to those skilled in the art and such approaches otherthan those described herein are included in the instant invention. In apreferred embodiment vegetable oil that contains biologically activecompounds is treated with water and a suitable catalyst. The ensuingreaction converts the triglyceride to the corresponding fatty acids.After the conversion the molecular weight of the fatty acid compounds issubstantially reduced while the biologically active components withhigher molecular weights are not similarly reduced in molecular mass.Distillation will selectively remove the fatty acid compounds and leavea unique residue of biologically active materials with higher molecularweights. While the use of distillation is preferred for separation ofthe fatty acid component of the reaction it is obvious to one skilled inthe art that other methods of separating molecules that differ in sizethat could be used to separate the fatty acids from the biologicallyactive fraction. These methods are included in the present invention.The products of the current invention may be produced using enzymatic,organic and mineral catalysts and as these catalysts are known to thoseskilled in the art of lipid chemistry they are included as a portion ofthe current art.

Molecular Weight Reduction: Saponification

In a preferred embodiment of the present invention the glyceridecomponent of the seed oil is converted to soaps which may be acidulatedto release fatty acids. There are many documented approaches to thechemical conversion of triglycerides to soaps known by those skilled inthe art and such approaches other than those described herein areincluded in the present invention. In a preferred embodiment vegetableoil that contains biologically active compounds is treated with waterand a suitable base. The ensuing reaction converts the triglyceride tothe corresponding soap. After the conversion the soaps may be convertedby the addition of a suitable acid to yield a solution of fatty acidsand the biologically active fraction. The molecular weight of the fattyacid compounds is substantially reduced while the biologically activecomponents with higher molecular weights are not similarly reduced inmolecular mass. Distillation will selectively remove the fatty acidcompounds and leave a unique residue of biologically active materialswith higher molecular weights. While the use of distillation ispreferred for separation of the fatty acid component of the reaction itis obvious to one skilled in the art that other methods of separatingmolecules that differ in size could be used to separate the fatty acidsfrom the biologically active fraction. These methods are included in theinstant invention. The products of the current invention may be producedusing a wide range of alkali materials known to those skilled in the artof lipid chemistry; the use of these materials is included as a portionof the current art.

Molecular Weight Reduction: Reduction

In a preferred embodiment of the current invention the glyceridecomponent of the seed oil is converted to fatty alcohols. There are manydocumented approaches to the chemical conversion of triglycerides tofatty alcohols known by those skilled in the art and such approachesother than those described herein are included in the instant invention.In a preferred embodiment vegetable oil that contains biologicallyactive compounds is treated with metallic potassium in butanol. Theensuing reaction converts the triglyceride to the corresponding alkanol.The molecular weight of the fatty alcohol compounds is substantiallyreduced while the biologically active components with higher molecularweights are not similarly reduced in molecular mass. Distillation willselectively remove the fatty alcohol compounds and leave a uniqueresidue of biologically active materials with higher molecular weights.While the use of distillation is preferred for separation of the fattyalcohol component of the reaction it is obvious to one skilled in theart that other methods of separating molecules that differ in size couldbe used to separate the fatty alcohols from the biologically activefraction. These methods are included in the present invention. Theproducts of the current invention may be produced using other alkalimetals and by other reactions known to those skilled in the art of lipidchemistry; the use of these reactants and catalysts is included in thepresent invention.

Distillation

Wide ranges of distillation processes are known to those skilled in theart of lipid chemistry. It is known that lipid molecules are sensitiveto damage by exposure to high temperatures encountered in distillationand as such distillation processes that minimise temperature exposureare preferred. Vacuum speeds distillation and minimises exposure toheat. Stills that operate under vacuum are thus preferred. Examples ofpreferred processes also include continuous distillation methodsincluding but not limited to molecular distillation, thin filmdistillation and other short path and continuous distillation processes.

Size Exclusion Chromatography

It is also possible to separate compounds utilising size exclusionchromatography. In a preferred method higher molecular weightbiologically active compounds are separated from lower molecular weightfatty compounds by passage over suitable size exclusion media. Examplesof suitable media include but are not restricted to Sephadex LH-20 andStyragel GPC.

Measurement of Carotenoid

Carotenoids can be measured in whole vegetable oil and in concentratesby the presence of specific peaks in the visible range of the spectrumusing a suitable spectrophotometer. The carotenoid content can beestimated utilising a standard curve prepared from a pure standard.Carotenoids were estimated on the basis of either beta carotene orlutein standards.

Measurement of Sterol

Sterol content was determined by non-destructive NMR analysis. In thisprocedure the oil or biologically active concentrate was dissolved indeuterated chloroform and the proton spectrum was recorded using a 400MHz Bruker Spectrospin spectrometry. Based on standard curvesestablished on solutions of phytosterol free esters and cholesterol itwas determined that spectrometry could reliably determine theconcentration of sterols in vegetable oil samples.

GC-FID and GC-MS was used to determine sterol concentration in fattyacids and esters.

Measurement of Tocopherol

GC-FID and GC-MS was used to determine tocopherol concentration in fattyacids and esters.

Measurement of Squalene

GC-FID and GC-MS was used to determine squalene concentration in fattyacids and esters.

Measurement of Dolichol

LC-MS was used to determine the presence of dolichol in fatty acid,ester and

Lubricity Measurements:

Laboratory Method:

Lubricity is measured using a Munson Roller On Cylinder LubricityEvaluator (M-ROCLE; Munson, J. W., Hertz, P. B., Dalai, A. K. andReaney, M. J. T. Lubricity survey of low-level biodiesel fuel additivesusing the “Munson ROCLE” bench test, SAE paper 1999-01-3590). TheM-ROCLE test apparatus conditions are given in Tablel. During the test,the reaction torque was proportional to the friction force produced bythe rubbing surfaces and was recorded by a computer data acquisitionsystem. The recorded reaction torque was used to calculate thecoefficient of friction with the test fuel. The image of each wear scarproduced on the test roller was captured by a video camera mounted on amicroscope and was transferred to image processing software, from whichthe wear scar area was measured. After determining the unlubricatedHertzian contact stress, a dimensionless lubricity number (LN),indicating the lubricating property of the test fuel, was determinedusing the following equation:LN=●_(ss)/●_(H)●_(ss); ●_(ss)=P/AWhere:

-   ●_(ss)=steady state ROCLE contact stress (mPa);-   ●_(●) ● Hertzian theoretical elastic contact stress (mPa);

Kerosene Reference Fuel was Escort Brand 1-K Triple Filtered, LowSulfur, Canadian Tire Stock No. 76-2141-2, Lot 135, BO2943. Each fuelester sample was lubricity tested six times on the machine followed by acalibration of the reaction torque. TABLE 1 M-ROCLE TEST CONDITIONS Fueltemperature, ° C. 25 ± 1.5 Fuel capacity, mL 63 Ambient temperature, °C. 24 ± 1.0 Ambient humidity, % 35-45 Applied load, N 24.6 Loadapplication velocity, mm/s 0.25 Test duration, min 3 Race rotationalvelocity, rpm 600 Race Surface velocity, m/s 1.10 Test specimens Falextest cylinder, F-S25 test rings, SAE 4620 steel Outer diameter, mm 35.0Width, mm 8.5 Falex tapered test rollers, F-15500, SAE 4719 steel Outerdiameter, mm 10.18, 10.74 Width, mm 14.80Field Test Method:

Motor oil analysis was utilized to infer engine wear. This involvedhigh-resolution Inductively Coupled Plasma (ICP) Spectrometry analysisof the used oil wear particles and oil additive elements. Ferrography,and magnetic particle analysis was determined for larger (>5 μm) wearparticles. Physical and chemical analyses of oil viscosity, acidneutralizing-ability (Total Base Number (TBN) and Total Acid Number(TAN)), and any dilution by fuel, water, or glycol was also monitored.An independent laboratory, Fluid Life Corporation in Edmonton Alberta,conducted these analytical tests.

All motor oil analysis data was adjusted to calculate true wear ratesconsidering oil volumes present in the crankcase, oil consumed, samplevolumes, and oil additions. All wear metals were monitored, with enginewear iron examined most critically. As well, by sectioning the filtersafter each oil change, filter wear and contaminant particles weremicroscopically and spectrographically compared. Field test logsindicating daily ambient minimum and maximum temperatures, numbers ofcold and hot starts, ratios of city to highway driving, and liters offuel consumed were tabulated. Consistent driving styles were enforced.Fuel economy and any operational difficulties were noted throughout thetest program. Esso brand regular unleaded gasoline and PennzoilMultigrade SJ motor oils were used throughout the study. The canolaadditives were prepared or obtained as described in specific examples.

Calculation of True Wear Rate

Consider for example, a vehicle engine that operates “normally” or“ideally”, generating and depositing in the crankcase oil a constant 10parts per million (10 ppm) of iron (Fe) in every 1,000 km of operation.Its “true wear rate” would be calculated by dividing the particle countby the distance traveled, yielding 10 ppm/1,000 km. Here, round numbershave been used to assist the reader in understanding the procedure. Ifthe vehicle were operated for 10,000 km under uniform conditions thewear iron level would rise 10 fold to 100 ppm Fe. This rise in ppm couldstart from zero ppm for an initially “flushed clean” engine, or moreoften from some initial “reference” level, taken shortly after an oilchange. A typical oil and filter change typically leaves 10% to 15% ofthe used oil behind, so referencing is an important initial first stepin a comparative engine wear analysis.

If the crankcase capacity of the example engine is 10 L, the amount ofelemental iron deposited in the oil after 10,000 km can be calculated asfollows:

The 100 ppm Fe is present in the 10 L crankcase volume.

Therefore the iron wear volume is obtained by multiplying the ironconcentration by the oil volume:100 parts Fe(10⁻⁶)×10 L=1,000 μL Fe.

This 1,000 μL Fe is the engine wear volume under ideal 10,000 kmconditions.

If the engine oil was referenced at, say 70 km, and found to contain 10ppm Fe, this would cause the final test reading after the 10,000 km tobe 10 ppm higher:100 ppm+10 ppm=110 ppm.

So to correct for initial residual iron one must subtract the referenceppm from the final test ppm, to obtain the “net” wear iron, which inthis case is still:110 ppm−10 ppm=100 ppm.

Oil sampling itself requires a small amount of oil (˜200 mL) to bewithdrawn from the crankcase each time the wear metals are monitored.

Assume 5 oil samples of 0.2 L=1.0 L of oil was removed during the 10,000km run. The average net ppm Fe concentrations in these 5 samples wouldbe close to the average net crankcase concentration of 50 ppm, whichstarted at 0 ppm and ended at 100 ppm.

This oil sampling has caused two things to happen:

-   (a) There is now 1.0 L less oil in the 10.0 L crankcase due to the    sampling, i.e. 9.0 L.-   (b) 1.0 L of oil containing, on net average, ˜50 ppm Fe has been    removed.

The indicated final net test value would no longer equal 100 ppm Fe butcan be calculated by doing a wear iron balance on the removal of ironactivity as follows:(100 ppm×10 L)−(50 ppm×1 L)=Test Fe ppm×9 L,

Solving for the Test Iron level in ppm, we obtain:Test ppm=(1000 μL Fe−50 μL Fe)/9 L,Test ppm=950 μL/9L=105.5 ppm Fe.

Due to sampling the “wear rate” based on the final test value of 105.5ppm Fe, instead of the true net previous 100.0 ppm value, would becalculated in error as too high at:105.5 ppm Fe/10,000 km, or, 10.55 ppm Fe/1000 km.

To compensate for sampling, “adding back” the oil sample volumes withnew oil, each time a sample was taken, could be tried. New oil maycontain small levels of wear metals (0.0-2.0 ppm Fe) and high levels ofadditive metals (800-1200 ppm Zn).

Focusing, for now, on the iron, we can do another iron balance takinginto account the 1.0 L sampling volumes and the 1.0 L add-back volumes(at 1 ppm Fe for new oil) as follows, starting with the previous truewear iron level:(100 ppm×10 L)−(50 ppm×1 L)+(1 ppm×1L)=Test ppm×10 L  (Eq. 1)Test ppm=(1000 μL Fe−50 μL Fe+1 μL)/10 LTest ppm=951/10=95.1 ppm FeAfter taking samples, and adding oil back, the indicated wear rateresult based on the final sample is now too low, at 95.1 ppm Fe/10,000km or 9.51 ppm Fe/1000 km.

If an engine “uses” oil, this volume will be similar to us taking outoil samples. If the oil is “topped-up” to the full mark, this is likeadding back new oil after sampling. If the crankcase ends up below orabove “full”, this can also be taken into account with reference to theprevious two examples.

It is desired to calculate the “true ppm” based on a “test ppm” wearindication. In more general terms the previous iron balance (Eq. 1) canbe rewritten as follows:(True ppm×Start L)−(True ppm×Used L/2)+(New ppm×Add L)=Test ppm×Test L$\begin{matrix}{{{True}\quad{ppm}} = \frac{\begin{matrix}{\left( {{Test}\quad{ppm} \times {Test}\quad L} \right) +} \\{\left( {{True}\quad{ppm} \times {Used}\quad{L/2}} \right) - \left( {{New}\quad{ppm} \times {Add}\quad L} \right) -}\end{matrix}}{{Start}\quad L}} & \left( {{Eq}.\quad 2} \right)\end{matrix}$For True ppm, we can approximate the True ppm in the second term of (Eq.2) equal the Test ppm, to get (Eq. 3): $\begin{matrix}{{{True}\quad{ppm}} = \frac{\begin{matrix}{\left( {{Test}\quad{ppm} \times {Test}\quad L} \right) +} \\{\left( {{Test}\quad{ppm} \times {Used}\quad{L/2}} \right) - \left( {{New}\quad{ppm} \times {Add}\quad L} \right) -}\end{matrix}}{{Start}\quad L}} & \left( {{Eq}.\quad 3} \right)\end{matrix}$Using the Test 95.1 ppm value from the example above, and substitutinginto (Eq. 3), yields a reasonably good True Fe value, close to the known100.0 ppm, as: ${\underset{Fe}{True}\quad{ppm}} = {\frac{\begin{matrix}{\left( {95.1\quad{ppm} \times 10\quad L} \right) +} \\{\left( {95.1\quad{ppm} \times 1\quad{L/2}} \right) - \left( {1\quad{ppm} \times 1\quad L} \right)}\end{matrix}}{10\quad L} = {99.75\quad{ppm}}}$If a higher accuracy is required this 99.75 ppm value can be substitutedfor the Test ppm yielding:${\underset{Fe}{True}\quad{ppm}} = {\frac{\begin{matrix}{\left( {95.1\quad{ppm} \times 10\quad L} \right) +} \\{\left( {99.75\quad{ppm} \times 1\quad{L/2}} \right) - \left( {1\quad{ppm} \times 1\quad L} \right)}\end{matrix}}{10\quad L} = {99.99\quad{ppm}}}$Therefore the following, repeated, Equation 3 can be used to calculate“True Wear” or “Normalize” indicated lubricant test results based on oilvolumes used or sampled, crankcase capacity, new oil added, or anycombination of the above: $\begin{matrix}{{True}\quad{ppm}\frac{\begin{matrix}{\left( {{Test}\quad{ppm} \times {Test}\quad L} \right) +} \\{\left( {{Test}\quad{ppm} \times {Used}\quad{L/2}} \right) - \left( {{New}\quad{ppm} \times {Add}\quad L} \right) -}\end{matrix}}{{Start}\quad L}} & \left( {{Eq}.\quad 3} \right)\end{matrix}$

EXAMPLES Example 1 Two Stage Transesterification of Canola Oil withMethanol and Potassium Hydroxide

Methyl esters of canola oil, also known to those skilled in the art aslow erucic acid rapeseed oil, were prepared using a two-stage basecatalysed transesterification. The two-stage reaction was required toremove glyceride from the final product. Prior to the reaction thecatalyst was prepared by dissolving potassium hydroxide (10 g) inmethanol (100 g). The catalyst solution was divided into two 55 gfractions and one fraction was added to 500 g of canola oil (purchasedfrom a local grocery store) in a 1 L beaker. The oil, catalyst andmethanol were covered and stirred vigorously for 1 hour on a stirringhot plate by the addition of a teflon stirring bar. After stirring, thecontents of the beaker were allowed to settle for 2 hours. At this timea cloudy upper layer and a viscous lower layer had separated. The layerswere separated using a seperatory funnel and the upper layer was mixedwith the remaining potassium hydroxide in methanol solution. This secondmixture was stirred vigorously in a covered beaker for 1 hour andallowed to settle overnight. The mixture settled to form two layers. Theupper layer was collected using a seperatory funnel and used for furtherrefining steps.

Example 2: Two Stage Transesterification of Tallow with Methanol andPotassium Hydroxide

Tallow was collected from a renderer. Five hundred grams of tallow wereheated to 40° C. prior to esterification to liquify the solid mass.Thereafter, all processes and conditions were identical to thosedescribed in example 1.

Example 3: Refining and Distillation of Canola Oil Methyl Ester

Canola methyl ester prepared in example 1 was refined to removemethanol, glycerol, soaps and other compounds that might interfere withdistillation. Methanol was removed under vacuum (28.5″) by a rotaryvacuum evaporator equipped with a condenser. The methyl esters weremaintained at 50° C. for 30 minutes to thoroughly remove alcohol. Afterevaporation the esters were treated with silica (0.25% w/w Trisyl 600;W. R. Grace Co.) and stirred at room temperature for 1 hour. Aftersilica treatment methyl esters were filtered over a bed of Celite toremove both silica and other materials.

After refining the methyl esters, fractional high vacuum distillationwas performed using a simple distillation apparatus. A vacuum of lessthan 1 mm was maintained throughout the procedure. During fractionationtemperatures at the top of the column, before the condenser, werebetween 120° C. and 140° C. The distillation apparatus included a liquidnitrogen cooled vapour trap, which allowed the attainment of high vacuumconditions. Approximately 500 mL of distillate (about half the sample)was obtained and then the heating mantle was removed while maintainingthe apparatus under vacuum. Vacuum was then broken and fractions of bothdistillate and bottoms were obtained for further studies. Distillationwas then resumed until a further 200 mL of distillate were obtained(about half the sample). The apparatus was again chilled, vacuum wasbroken and samples of 100 mL of both bottoms and distillate wererecovered. All samples of bottoms and distillate were analysed todetermine the content of soaps and free fatty acids using AOCS methodsCc 17-95 and Ca 5a-40 respectively.

Some samples of column bottoms were noted to have elevated levels offree fatty acids. These samples were treated by briefly contacting witha mixture of 1 molar potassium hydroxide dissolved in glycerol toconvert the fatty acids to soaps. The glycerol phase was easilyseparated from the oil phase by decanting. Following alkaline glyceroltreatment silica (0.25% w/w Trisyl 600) and was added to the oil phaseand the phase was filtered over a bed of celite.

Example 4: Refining and Distillation of Tallow Methyl Ester

Tallow esters were refined and distilled as described for rapeseedesters in Example 3.

Example 5: Lubricity Testing of Methyl Canola and Tallow Esters

Lubricity was measured using a Munson Roller On Cylinder LubricityEvaluator (M-ROCLE; Munson, J. W., Hertz, P. B., Dalai, A. K. andReaney, M. J. T. Lubricity survey of low-level biodiesel fuel additivesusing the “Munson ROCLE” bench test, SAE paper 1999-01-3590). TheM-ROCLE test apparatus conditions are given in Table1. M-ROCLE operationand equations used to describe lubricity number are described above.Table 2 describes the samples subjected to analysis.

Lubricity testing was performed on the first distillate and columnbottoms, which constituted about a four-fold concentrate of high boilingsubstances. A total of 6 replications were performed to allow forstatistical analysis. All tests were performed on a 1% solution ofconcentrate or distillate in kerosene. Table 3 contains the results ofanalyses.

In testing it was found that kerosene produced the lowest lubricitynumber and that all treatments increased lubricity number with respectto controls. Among the treated samples the concentrates consistentlydemonstrated the highest lubricity numbers. The lubricity numbers forconcentrates of canola and the two tallow samples were not significantlydifferent from each other and in all cases the concentrates had greaterlubricity than the distillates. The lubricity numbers noted for thedistillates were lower than the concentrates, though higher thancontrols, indicating that only half of the improvement in lubricitynumber was contributed by the distilled methyl ester. In the two tallowsamples it was found that prior to distillation the lubricity number wassimilar to the lubricity number for the concentrate.

Uniformly it was found that all treatments also decreased wear scararea. Surprisingly it was found that although distilled methyl esterssignificantly decreased wear scar area concentrates produced the lowestwear scar areas. For example, tallow 1 methyl ester (sample number 4)produced a wear scar area of 0.2410 mm² while the distillate andconcentrate of this sample produced wear scars of 0.2763 mm² and 0.2446mm² respectively (Table 3).

It was discovered that the treatments had little impact on thecoefficient of friction in the current test. TABLE 2 Description ofrefining and distillation conditions used to prepare lubricity enhancedconcentrates All additive samples were Trisyl treated and CeliteFiltered Methyl Esters Bottle Base Material Fatty Bottle Sample # forMethyl Ester Acid % Wt. gr. #1 Canola Oil 0.04% 104 #2 Canola Oil 0.07%105 Distillate #3 Canola Oil 0.07% 84 Concentrate #4 Tallow 1 0.07% 93#5 Tallow 1 0.07% 96 Distillate #6 Tallow 1 0.10% 90 Concentrate #7Tallow 2 0.03% 88 #8 Tallow 2 0.06% 84 Distillate #9 Tallow 2 0.07% 98Concentrate

TABLE 3 Wear Scar Lubricity Area Standard Coefficient Sample NumberStandard (mm{circumflex over ( )}2) Deviation of Friction Standardnumber* (n = 6) Deviation (n = 6) [mm{circumflex over ( )}2] (n = 6)Deviation Kerosene 0.7547 0.0778 0.3195 0.0238 0.1142 0.0050 #1 0.86200.0579 0.2907 0.0029 0.1210 0.0034 #2 0.8341 0.0484 0.2783 0.0183 0.10950.0017 #3 0.9464 0.0706 0.2557 0.0121 0.1180 0.0022 #4 0.9561 0.05520.2410 0.0222 0.1136 0.0022 #5 0.8373 0.0352 0.2763 0.0120 0.1189 0.0020#6 0.9625 0.0456 0.2446 0.0102 0.1183 0.0019 #7 0.9348 0.0438 0.26230.0113 0.1163 0.0023 #8 0.8513 0.0492 0.2723 0.0092 0.1116 0.0013 #90.9555 0.0712 0.2547 0.0162 0.1182 0.0009*number corresponds to sample number in table 2

Example 6 Impact of Oil Extraction and Refining Procedures on theLubricity of Canola Oil

Approximately twenty kg (20.8) of canola seed was crushed in a Kometexpeller press through a 6 mm die face producing 7.9 kg of oil withfines and 12.8 kg of meal. The oil was clarified by passing over glasswool followed by centrifugation at 2000×g for 15 min in a swing outrotor. The mass of the clarified oil was 7.2 kg. This oil was identifiedas pressed and unrefined or P-0. The meal arising from pressing wasextracted with hexane in 1.4 kg batches in a soxhlet extractor. Thehexane was collected and evaporated in a rotary evaporator producing 1.5kg of solvent extracted oil. This oil is identified as solvent extractedand unrefined or S-0. The combined oil yield from the two processes was42% of the original seed mass. The two samples of oil were used forfurther processing and analysis. Blending the crushed and solventextracted oils at a ratio of 5:1 produced the third sample. This oil isidentified as pressed, solvent extracted and unrefined or PS-0.

All oil samples were analyzed to determine the level of sterols (NMR),free fatty acids (AOCS Ca 5a-40), minerals (ICP) and lubricity (MunsonROCLE).

Oils (P-0, S-0 and PS-0) were degummed by adding 0.2% by weight of fiftypercent citric acid to the oil while heating to 40-45° C. for 30 minuteswith agitation. After reaction with the acid an additional of 2% ofwater (w/w) was added. The water treated oils were then heated to 60-70°C. for a further 20 minutes then centrifuged (2,000×g for 15 minutes).The upper layer of clear oil was recovered and analyzed to determineFFA, minerals and lubricity. Degumming produced three oil products:pressed degummed oil, P-1; solvent extracted degummed oil, S-1; andpressed and solvent extracted degummed oil PS-1

Approximately 300 g of each oil (P-1, S-1 and PS-1) was neutralized oralkali refined, for further analyses and processing. Alkali refining wasachieved by adding a solution of 10% (w/w) sodium hydroxide to thedegummed oil. The free fatty acid level was used to determine thestoichiometric amount of sodium hydroxide solution required forneutralization with a small excess. Neutralization was accomplished at60-70° C. with a reaction time of 5 minutes with agitation. Afterneutralization the oil and soap water solution were separated bycentrifugation (2,000×g for 15 minutes). The oil had a cloudyappearance. Evaporation of the cloudy oil produced clear oil that wasanalyzed for FFA, minerals and lubricity. Neutralization produced threeoil products: Pressed neutralized oil, P-2; solvent extractedneutralized oil, S-2; and pressed and solvent extracted neutralized oilPS-2.

The alkali refined, neutralized oils (P-2, S-2 and PS-2) were bleachedby the addition of 1% (w/w) bleaching clay to oil that had beenpreheated to 110° C. under vacuum. The oil was agitated in the presenceof the bleaching clay for 30 min after which the temperature was allowedto fall to 60° C. prior to release of the vacuum. The oil and clay werethen filtered through a bed of celite and Whatman No. 1 filter paper ina Buchner funnel. The filtered oil was analyzed to determine FFA,minerals and lubricity. Bleaching produced three oil products: Pressedbleached oil, P-3; solvent extracted bleached oil, S-3; and pressed andsolvent extracted bleached oil PS-3.

In the final stage of processing the oils (P-3, S-3 and PS-3) weredeodorized by passage through a 2.0 inch diameter Pope wiped film still.The still was adjusted to deliver oil at 2 mL/min, evaporationtemperature was 170° C. and vacuum was 10⁻² mbar. Deodorizing producedthree oil products: Pressed deodorized oil, P-4; solvent extracteddeodorized oil, S-3; and pressed and solvent extracted deodorized oilPS-3.

Sterol is observed as a peak at 0.66 ppm in the proton spectrum. Thepeak is small but may be quantified with a sufficiently powerfulspectrometer. The level of sterol in the solvent extracted portion ofthe oil is approximately the level found in the pressed oil (Table 4).With the exception of deodorizing treatments none of the refining stepsaffected the measured level of sterol.

Nine different mineral elements are observed in the ICP data includingsilicon, sodium, potassium, iron, boron, phosphorous, zinc, calcium, andmagnesium. The amounts of most minerals are higher in solvent extractedoils than the pressed oil. Refining tends to remove minerals but itseffect is different among the three samples. Degumming reduced thephosphorous content of pressed oil from 8 to 4 ppm (P-0 vs P-1) and from168 to 57 ppm in the mixed oil (PS-0 vs. PS-1) but had no effect on thelevel of phosphorous (1030 ppm) in the solvent extracted oil (S-0 vs.S-1). Upon completion of all refining steps the pressed oil wasvirtually devoid of all mineral contamination showing only traces of tin(1 ppm, probably spurious) and silicon (7 ppm). Refining similarlyimproved the quality of the mixed oil (PS-4) where only traces ofsilicon, phosphorous, calcium and magnesium (3,2,2 and 2 ppmrespectively) were observed. Full refining was not useful in removingmaterials from the solvent extracted oil where silicon, sodium,phosphorous, calcium and magnesium were observed at appreciable levels(10, 41, 197, 225 and 69 ppm respectively). Trace levels of potassiumand lead were reported but the latter measurement was likely spuriousinstrument noise.

The effect of the three oils at all stages of refining on kerosenelubricity was evaluated by preparing a 1% (w/w) solution in kerosene andtesting in a Munson Roller On Cylinder Lubricity Evaluator to determinethe coefficient of friction and wear scar area. Lubricity number (LN)was calculated from the two numbers. Wear scar area was greatly reducedby all treatments. Several differences were observed among treatmentsbut generally the size of differences among treatments was much smallerthan the difference between untreated kerosene and the individualtreatments. Wear scar area was for all three unrefined oils from alltreatments. Degumming resulted in oils that produced a larger wear scar.Other refining treatments did not affect wear scar significantly.

All treatments lowered the coefficient of friction but substantialdifferences among treatments were observed. Alkali refined oils that hada greater coefficient of friction in all cases while bleaching reducedfriction coefficients only for solvent extracted oil (S and PS, Table4). Deodorizing also increased the coefficient of friction for the twosolvent extracted oils. On average the coefficient of friction waslowest in oils containing the solvent extracted components.

Lubricity number reflects the effect of the oil on both wear scar andcoefficient of friction. All oils regardless of the treatment increasethe lubricity number. The solvent extracted oil provided the greatestincrease in lubricity number over the blended and pressed oil types.Refining does not appear to affect the LN of pressed oil while it doesresult in interesting changes in the LN of the solvent extractedfractions. In the solvent extracted oils it is seen that degumming theoil lowers LN. Alkali refining has little additional affect on LN butbleaching appears to restore the LN though not to the levels observed inunrefined oil. Deodorizing lowers LN in the solvent extracted and theblend oils. TABLE 4 Effect of oil refining on select metal componentconcentrations and lubricity factors wear Si Na K B P Zn Ca Mg Sterolscar FFA (%) (PPM) (PPM) (PPM) (PPM) (PPM) (PPM) (PPM) (PPM) (NMR) (•M²)C of F* LN P**-0*** 1.244 0 0 1 1 8 1 12 3 0.024 0.2634 0.1270 0.8193P-1 1.231 1 1 0 3 4 0 1 1 0.021 0.2732 0.1179 0.8507 P-2 0.084 1 7 0 2 10 0 0 0.022 0.2830 0.1239 0.7800 P-3 0.070 1 0 0 1 0 0 0 0 0.021 0.26890.1222 0.8359 P-4 0.056 7 0 0 0 0 0 0 0 0.018 0.2754 0.1218 0.8167 PS-01.866 2 1 32 1 168 1 70 33 0.240 0.2519 0.1143 0.9543 PS-1 1.840 2 1 8 257 0 20 9 0.011 0.2944 0.1092 0.8527 PS-2 0.141 1 2 0 1 5 0 4 0 0.0270.2877 0.1233 0.7722 PS-3 0.126 1 0 0 0 3 0 2 1 0.023 0.2716 0.11430.8844 PS-4 0.084 3 0 0 0 2 0 2 2 0.007 0.2870 0.1171 0.8146 S-0 4.57310 8 209 1 1030 3 368 190 0.040 0.2365 0.1127 1.0318 S-1 5.434 12 10 2073 1040 3 378 190 0.042 0.2658 0.1143 0.9006 S-2 0.310 10 45 4 1 207 0273 74 0.034 0.2504 0.1228 0.8960 S-3 0.364 10 42 3 1 199 0 255 71 0.0350.2601 0.1082 0.9738 S-4 0.364 10 41 3 0 197 0 255 69 0.033 0.25780.1241 0.8559*Coefficient of friction**P = pressed oil, PS = pressed and solvent extracted oil S = solventextracted oil***0 = unrefined, 1 = Degummed, 2 = Degummed and neutralized, 3 =Degummed, neutralized and bleached, 4 = Degummed, neutralized, bleachedand deodorized.

Example 7: Influence of Canola Oil Additization on Wear and Fuel Economy

This example describes the canola lubricity field performance of a fullywear documented gasoline engine, a 3.0 L V6 Toyota Camry. Tests beganwith an additization rate of 250 ppm Canola Oil in unleaded commercialgasoline under summer driving conditions. To reference these tests acontrol summer test of 10,000 km was conducted without the canola oilpresent. The same motor oil Pennzoil SJ SAE 10W-30 was used through outthe reference and treatment test periods. Eight oil samples were taken.Data was analyzed in two parts, 0 to 5,800 km and 5,800 km to 10,510 km.The driving was 65% highway and 35% city. Starts totaled 458 Cold and327 Hot. Ambient temperatures ranged from a mean minimum of 8.5° C. to amaximum of 20.8° C. Table 5 shows the comparison of net wear iron ppmlevels generated up to the 6,000 summer distances with regular gasolineand with 250 ppm canola oil additization.

Canola oil supplemented gasoline produced a significant ICP wearreduction compared with the control. The overall averaged wear rate withregular gasoline was 0.99 ppm Fe/1,000 km while the instantaneous methodyielded a rate of 0.87 ppm Fe/1,000 km for the reference fuel. Thereference results exceeded the 0.63-0.66 ppm Fe/1,000 km obtained withcanola oil present and revealed that canola oil additized fuel hadresulted in a 33% wear reduction overall and a 26% reductioninstantaneously. The average mileage obtained with canola oil presentwas 28.1 MPG while reference gas mileage was 4% better at 29.3 MPG. Inthis test canola oil additization lowered fuel economy.

In Table 6 the ferrography for reference gasoline revealed a wearparticle density of 15 with other contaminants counting 8. The canolaoil additized fuel run analysis indicated 14 for wear particles and 8for other debris, indicating no effect of the treated fuel on largerferrographic particles.

The filter analysis with 250 ppm canola oil additized fuel reveals rust,dirt, and varnish particles. The largest translucent particles ofvarnish measure about 200 μm. The spectrographic analysis of the filterresidues indicated silicon, iron, copper traces and sodium. The presenceand level of the contaminants is normal.

Both neutralization numbers were not affected significantly by canolaoil treatment. Motor oil taken from the vehicle after operation on 250ppm canola oil additized fuel lowered the total base number to 6.06while the total acid number remained at 3.66 (Table 6).

After summer operation on gasoline containing 250 ppm canola oil (6,261km) viscosity was lowered to 57.6 cSt at 40° C. and 8.95 cSt at 100° C.This represented a 17% drop in viscosity at 40° C. and an 18% change at100° C. Also the presence of 1% fuel dilution of the oil was indicatedafter driving 10,243 km, when the oil was changed.

Example 8: Influence of Canola Methyl Ester Additization on Wear andFuel Economy

This example describes the Canola lubricity field performance of a fullywear documented gasoline engine, a 3.0L V6 Toyota Camry. Tests beganwith an additization rate of 125 ppm canola oil methyl ester (CME) inunleaded commercial gasoline under summer driving conditions. Toreference these tests a control summer test of 10,000 km was conductedwithout the canola methyl ester present. The same motor oil Pennzoil SJSAE 10W-30 was used through out the reference and treatment testperiods. For canola methyl ester additization tests a distance of 10,017km was covered with 74% highway driving. Cold starts added up to 278while hot starts equaled 311. Temperature means ranged from 12.3°C. to25.4° C.

The ICP iron wear rates were remarkably low with the 125 ppm CMEtreatment (Table 5). The overall rate method yielded only 0.50 ppmFe/1,000 km while the instant point-to-point mean was similar at 0.48ppm Fe/1,000 km. This lower CME treatment resulted in 49% to 45% wearreduction compared to the unadditized reference. It is clearlyillustrated that CME wear performance is superior to both the referenceand the 250 ppm canola oil additized fuel performance. Both canolaadditives are considerably better than the reference regular gasoline.The calculated mean fuel economy with 125 ppm CME was some 5% betterthan for the reference gasoline, yielding 30.8 MPG compared to theformer 29.3 miles per Imperial gallon on regular gasoline.

The consistency of the reference wear readings were established bycomparing average ICP data wear rates (Table 5) for regular gasoline.These averages were 0.87, 0.85, 0.99 and 0.87 ppm Fe/1,000 km. On thebasis of this long-term reference, the listed per-cent summer wear ratereductions were 33% and 28% for instantaneous and cumulative wear whenoperating on 125 ppm CME.

Ferrography analysis of motor oil obtained after operation on 125 ppmCME totaled 6 wear particles and 2 other particles. This represents areduction of 60% and 87% reduction from reference analysis. Most ofthese wear metals were described in the ferrography reports as “lowalloy steel showing rubbing/sliding wear” although it is difficult todistinguish between very small steel and cast-iron particles,originating from the cylinder block.

The last filter obtained after operation on 125 ppm CME had far lessdebris in it compared to the other two filters. The white filter papersupport shows through the particles, which are at a much lowerconcentration. Dirt/dust, rust and varnish are the major contaminants.The presence of silicon, iron, and traces of lead, copper and tinappeared spectrographically.

Operation on the CME additized fuel lowered the TBN to 6.19 while theTAN climbed to 4.20. This revealed that both neutralization numbers werenot affected significantly by the Canola methyl ester.

Viscosity of the motor oil was also determined after operation on 125ppm CME. After the 10,016 km ended, the oil tested 59.4 at 40° C., a 13%drop. For 100° C. the values 9.43 cSt were reported, with a 14% drop.Viscosity performance was within specifications

With 125 ppm Canola Methyl Ester added to the gasoline engine wear ratewas reduced by almost one-half, to only 0.5 ppm Fe/1,000 km, potentiallydoubling engine life. Field fuel economy rose by 5%. The engine oilremained within neutralization and viscosity specifications after some10,000 km of field-testing. The ferrographic and oil filter debrislevels were markedly reduced and appeared normal. Furthermore nodriveability or other engine performance problems were detected as theresult of the specific CME treatment rate used in unleaded regulargasoline.

Example 9: Winter Canola Oil Gasoline Field Testing, Wear and FuelEconomy

This example describes the Canola lubricity field performance of a fullywear documented gasoline engine, a 3.0 L V6 Toyota Camry. Tests beganwith an additization rate of 250 ppm canola oil in unleaded commercialgasoline under winter driving conditions. To reference these tests aseries of winter reference runs were performed without the additive. Thesame motor oil Pennzoil SJ SAE 10W-30 was used through out the referenceand treatment test periods.

The reference wear rate data is recorded in Table 5 reflecting theaccumulation of iron (ppmFe/1,000 km value) averaged 2.24 (overall) and1.91 (measuring point to point). Reference gasoline economy recordsaveraged 24.5 MPG. The numbers of cold and hot starts during the winterreference period were recorded. Mean ambient winter temperatures were inthe −15° C. to −7° C. range. The proportion of highway driving wascalculated as 71% and 43% for the reference tests.

The canola oil additive was pre-mixed with 50% gasoline to facilitatetank blending upon cold refueling. The canola oil test data involved 224cold and 101 hot starts with 72% highway driving. The fuel economy roseto 27.5 MPG, a 12% improvement in referenced shorter-term mileage. Table5 compares regular gasoline and the 250 ppm canola oil additive.Calculations in Table 5 indicated that wear rates decreased slightlywith 250 ppm canola oil additized fuel, to 2.02 and 1.73 ppm Fe/1,000km. These reductions in wear were 6% and 20% based on the long-termreference and 10% and 9% based on the shorter-term comparative regulargas references.

For the canola oil additized fuel treatment, the level of ferrographicwear particles reached “12” while contaminants remained at “7”. Thisrepresented 11% lower wear particle count than previously referenced.The magnetic iron trend remained very low and unchanged at 0.2 μg/mL.

The oil filter taken after operation on 250 ppm canola oil additizedfuel revealed contaminants as dirt, rust and varnish. The spectrographicanalysis revealed iron, silicon, and traces of sodium, copper, andpotassium in the filter debris. Filter analysis results were normal.

The winter 250 ppm canola oil fuel additive resulted in a 5.8 TBN and a2.5 TAN indication. This 5.8 reading revealed a similar drop in reservealkalinity for TBN, noting the 5.7 TBN for the reference fuel. The TANof 2.5 for canola oil additized fuel treatment had not variedsignificantly from the 2.5 value for new oil or the 2.7 value for oilafter operation on the reference fuel.

Motor oil obtained after operation on 250 ppm canola oil additized fuelunder winter operation conditions had viscosity of 48.5 cSt at 40° C.and 8.73 cSt at 100° C. The viscosity had decreased 21% at 40° C. and17% drop at 100° C. from new oil. Compared to regular fuel, the relativeadditional loss of viscosity was 5% at 40° C. and 4% at 100° C. for thecanola oil additized gasoline.

The winter tests with 250 ppm canola methyl ester added to the gasolinewere encouraging. Engine wear rate was reduced by almost one-half, toonly 0.5 ppm Fe/1,000 km, potentially doubling engine life. Field fueleconomy rose by 5%. The engine oil remained within neutralization andviscosity specifications after some 10,000 km of field-testing. Theferrographic and oil filter debris levels were markedly reduced andappeared normal. Furthermore no driveability or other engine performanceproblems were detected as the result of the specific CME treatment rateused in unleaded regular gasoline.

Example 10: Winter Canola Methyl Ester Gasoline Field Testing, Wear andFuel Economy

This example describes the Canola lubricity field performance of a fullywear documented gasoline engine, a 3.0L V6 Toyota Camry. Tests beganwith an additization rate of 250 ppm canola methyl ester in unleadedcommercial gasoline under winter driving conditions. To reference thesetests a series of winter reference runs were performed without theadditive. The same motor oil Pennzoil SJ SAE 10W-30 was used through outthe reference and treatment test periods.

The reference wear rate data is recorded in Table 5 reflecting theaccumulation of iron (ppmFe/1,000 km value) averaged 2.24 (overall) and1.91 (measuring point to point). Reference gasoline economy recordsaveraged 24.5 MPG. The numbers of cold and hot starts during the winterreference period were recorded. Mean ambient winter temperatures were−7.9° C. and −3.7° C. the daily averaged minimum and maximums. Theproportion of highway driving was calculated as 71% and 43% for thereference tests.

The canola methyl ester tests spanned 4,202 km with 106 cold and 113 hotstarts logged with 72% highway driving. The average fuel economy duringthis test was 27.0 MPG, some 10% better compared to the regular gasreferences. Table 5 compares the net wear iron in the two winter testruns. The gasoline alone graph climbs higher than with 250 ppm thecanola methyl ester supplement. The engine-wear iron spectrometrycalculations revealed rates of 1.55 and 1.27 ppm Fe/1,000 km with canolamethyl ester. These were 28% and 41% lower than the long-term referencesand 31% and 41% below the shorter-term gasoline references as shown inTable 5. No driveability problems were experienced, with good power,starting, and stable idling rpm demonstrated while using 250 ppm canolamethyl ester as a gasoline additive.

With the canola methyl ester additive, ferrography indicated wearparticles were at the “13” level while a ranking of “8” appeared forcontaminants. Most metal particles are low alloy steel showingrubbing/sliding. Traces of copper/copper alloy (up to 40 microns)present were comments. The magnetic iron trend stayed minimally the sameat 0.2 μg/mL.

Analysis of the oil filter after operation on 250 ppm canola methylester in winter conditions indicated that contaminants were dirt, dust,rust and varnish. The debris texture looked fine with some metallicreflections. Spectrographic analysis revealed silicon, iron, and tracesof sodium, potassium, copper and tin in the residue. These filterresults were also judged normal.

Oil viscosity from oil taken after operation on canola methyl ester for4,104 km was 51.9 cSt at 40° C. and 9.46 at 100° C. No fuel dilution ofthe motor oil was observed during the trial. These test valuesrepresented similar viscosity to that obtained after similar operationon reference gasoline. The 250 ppm canola methyl ester treatment underwinter conditions appeared better in terms of viscosity dilution thanthe 250 ppm canola oil additive.

Example 11

Twenty liters of methyl esters were prepared according to example 1using canola oil obtained at a local grocery. The esters were thenplaced in 2 L lots in a high vacuum vessel used to feed a 2″ wiped filmevaporator (Pope Scientific, Saukville WI). Vacuum (0.01 torr) wasapplied to the high vacuum flask to remove residual volatile materials.After vigorous bubbling had ceased the material was passed through thewiped film still at an initial high rate (20 mL/min) to removelow-boiling materials. The walls of the still were heated to 80 C forthis process. During evaporation vapors were condensed by traps chilledwith liquid nitrogen. After removing removing volatiles from the methylester solution the still was heated to 170 C and the methyl esters werere introduced and the vacuum was maintained. The flow of liquid wasadjusted so that the flow of distillate was approximately 20 times theflow of residue. During this time 1.5 L of residue was collected. Theundistilled residue was introduced to the still and after distillationunder the same conditions a concentrate of 300 mL was obtained.

Analysis of the methyl esters and the canola oil with high field protonNMR Spectroscopy (500 MHz Bruker, Milton, ON Canada) revealed a smallbut observable peak in the spectrum contributed by plant sterols at 0.67ppm. The distillate did not have observable sterol peaks. Addition ofpure sterol (cholesterol or cholestanol) to the distillate restored thepeak. The residue of the distillation was also observed using high-fieldproton NMR. The proton spectrum was comparable to a mixture of plantfree and bound sterols with small amount of residual of methyl esters.With further preparation steps known to those skilled in the art, thefree and bound sterol fraction may be separated and used as componentsof nutritional concentrates.

Example 12 Production of Safflower Oil Ethyl Esters

Potassium hydroxide pellets (100 g) were dissolved in a 4 L beakercontaining 3500 g of absolute ethanol. The caustic ethanol solution wasadded to ten kg of safflower oil in a 20 L plastic pail held at roomtemperature and the mixture was stirred for 2 hours at room temperature.After 2 hours the solution was allowed to settle for 24 hours and theclear upper layer of ethyl esters was decanted into a clean plastic 20 Lpail. The lower layer was transferred to a 4 L separatory funnel and thelower layer of glycerin was separated from the remaining upper layer ofethyl esters. The recovered ethyl esters were combined with the decantedesters. The ethyl esters were then washed by the addition of 200 g ofwater and vigorous agitation of the solution. The water was allowed tosettle and the methyl ester layer was again decanted into a cleanplastic pail. The lower water layer was transferred to a 2 L separatoryfunnel and allowed to settle for 4 hours. The lower water layer wasdrained and the upper layer of washed methyl esters was combined withthe decanted washed esters. The washed esters were placed in a 20 Lrotary evaporator and all water and ethanol was removed by evaporationfor 2 hours at 80 C. The dried ester layer had a slightly cloudyappearance.

Celite (250 g) was mixed with a one liter portion of the cloudy esterlayer. The slurry was then used to form a filtration bed in a 20 cmclean and oven dry ceramic Buchner funnel. The first sample of ester wasreturned to the top of the filter bed. Thereafter the remaining volumeof ethyl esters was passed over the filter bed to remove particulatematter. Proton NMR and analysis of the fatty acid esters using gaschromatography indicated that the clear solution was greater than 95%fatty acid ethyl esters.

Example 13 Wiped Film Distillation of Safflower Oil Ethyl Esters

Ten liters of fatty acid ethyl esters were prepared according to example12 using safflower oil obtained at a local grocery. The esters were thenplaced in 2 L lots in a high vacuum vessel used to feed a 2″ wiped filmevaporator (Pope Scientific, Saukville Wis.). Vacuum (0.01 torr) wasapplied to the high vacuum flask to remove residual volatile materials.After vigorous bubbling had ceased the material was passed through thewiped film still at an initial high rate (20 mL/min) to removelow-boiling materials. The walls of the still were heated to 80 C forthis process. During evaporation vapors were condensed by traps chilledwith liquid nitrogen. After removing removing volatiles from the methylester solution the still was heated to 140 C and the ethyl esters werere introduced and the vacuum was maintained. The flow of liquid wasadjusted so that the flow of distillate was approximately 10 times theflow of residue. During this time 1 L of residue was collected. Theresidue of distillation was introduced to the still and afterdistillation under the same conditions a concentrate of 50 mL wasobtained.

Analysis of the ethyl esters and the safflower oil with high fieldproton NMR Spectroscopy (500 MHz Bruker, Milton, ON Canada) revealed twosmall but observable peaks in the spectrum contributed by plant sterolsand other triterpene alcohols. The distillate did not have observablesterol peaks. The residue of the distillation was also observed usinghigh-field proton NMR. The proton spectrum was comparable to a mixtureof ethyl esters with plant free and bound sterols and triterpenealcohols. With further preparation steps known to those skilled in theart the free and bound sterol fraction may be separated and used ascomponents of nutritional concentrates. The preparation may also be usedas a direct source of sterols.

Example 14: Two Stage Transesterification of Canola Oil with Methanoland Potassium Hydroxide

Methyl esters of canola oil, also known to those skilled in the art aslow erucic acid rapeseed oil, were prepared using a two-stage basecatalysed transesterification. The two-stage reaction was required toremove glyceride from the final product. Prior to the reaction thecatalyst was prepared by dissolving potassium hydroxide (190 g) inmethanol (3800 g). The catalyst solution was divided into two 1995 gfractions and one fraction was added to 20 L of canola oil (purchasedfrom a local grocery store) in a 30 L stainless steel pot. The oil,catalyst and methanol were covered and stirred vigorously for 1 hourwith an overhead stirrer. After stirring, the products of the reactionwere allowed to settle for 2 hours. At this time a cloudy upper layerand a viscous lower layer had separated. The majority of the upper layerwas decanted and the remaining layers were separated using a seperatoryfunnel. The upper layers were pooled, returned to the stainless pot withoverhead stirrer and the remaining potassium hydroxide in methanolsolution was added. This second mixture was stirred vigorously in acovered beaker for 1 hour and allowed to settle overnight. The mixturesettled to form two layers. The upper layer was collected by decantingand using a separatory funnel.

After separation of phases the upper layer was mixed with 400 mL ofwater. The water was removed from the upper phase by decanting. Thewashed esters were placed in a 20 L rotary evaporator and all water andethanol was removed by evaporation for 2 hours at 80 C. The resultingesters had a slightly cloudy appearance.

Celite (250 g) was mixed with a one liter portion of the cloudy esterlayer. The slurry was then used to form a filtration bed in a 20 cmclean and oven dry ceramic Buchner funnel. The first sample of ester wasreturned to the top of the filter bed. Thereafter the remaining volumeof methyl esters was passed over the filter bed to remove particulatematter. Proton NMR and analysis of the fatty acid esters using gaschromatography indicated that the clear solution was greater than 95%fatty acid methyl esters.

Example 15 Preparation of a Nutritional Concentrate From TransesterifiedCanola Oil and Analysis of a Potential Biologically Active Concentrate.

Twenty liters of methyl esters were prepared according to example 14using canola oil obtained at a local grocery. The esters were thenplaced in 2 L lots in a high vacuum vessel used to feed a 2″ wiped filmevaporator (Pope Scientific, Saukville Wis.). Vacuum (0.01 torr) wasapplied to the high vacuum flask to remove residual volatile materials.After vigorous bubbling had ceased the material was passed through thewiped film still at an initial high rate (20 mL/min) to removelow-boiling materials. The walls of the still were heated to 80 C forthis process. During evaporation vapors were condensed by traps chilledwith liquid nitrogen.

The still was then heated to 170 C and the methyl esters were reintroduced and the vacuum was maintained. The flow of liquid wasadjusted so that the flow of distillate was approximately 20 times theflow of residue. During this time 1.5 L of residue was collected. Theundistilled residue was introduced to the still and after distillationunder the same conditions a concentrate of 300 mL was obtained.

Analysis of the methyl esters and the canola oil with high field protonNMR Spectroscopy (500 MHz Bruker, Milton, ON Canada) revealed a smallbut observable peak in the spectrum contributed by plant sterols at 0.67ppm. The distillate did not have observable sterol peaks. Addition ofpure sterol (cholesterol or cholestanol) to the distillate restored thepeak. The residue of the distillation was also observed using high-fieldproton NMR. The proton spectrum was comparable to a mixture of of plantfree and bound sterols with small amount of residual of methyl esters.With further preparation steps known to those skilled in the art thefree and bound sterol fraction may be separated and used as componentsof nutritional concentrates.

Example 16 Transesterification of Safflower Oil with Ethanol.

Potassium hydroxide pellets (100 g) were dissolved in a 4 L beakercontaining 3500 g of absolute ethanol. The caustic ethanol solution wasadded to ten kg of safflower oil in a 20 L plastic pail held at roomtemperature and the mixture was stirred for 2 hours at room temperature.After 2 hours the solution was allowed to settle for 24 hours and theclear upper layer of ethyl esters was decanted into a clean plastic 20 Lpail. The lower layer was transferred to a 4 L separatory funnel and thelower layer of glycerin was separated from the remaining upper layer ofethyl esters. The recovered ethyl esters were combined with the decantedesters. The ethyl esters were then washed by the addition of 200 g ofwater and vigorous agitation of the solution. The water was allowed tosettle and the methyl ester layer was again decanted into a cleanplastic pail. The lower water layer was transferred to a 2 L separatoryfunnel and allowed to settle for 4 hours. The lower water layer wasdrained and the upper layer of washed methyl esters was combined withthe decanted washed esters. The washed esters were placed in a 20 Lrotary evaporator and all water and ethanol was removed by evaporationfor 2 hours at 80 C. The dried ester layer had a slightly cloudyappearance.

Celite (250 g) was mixed with a one liter portion of the cloudy esterlayer. The slurry was then used to form a filtration bed in a 20 cmclean and oven dry ceramic Buchner funnel. The first sample of ester wasreturned to the top of the filter bed. Thereafter the remaining volumeof ethyl esters was passed over the filter bed to remove particulatematter. Proton NMR and analysis of the fatty acid esters using gaschromatography indicated that the clear solution was greater than 95%fatty acid ethyl esters.

Example 17 Preparation of a Nutritional Concentrate from TransesterifiedSafflower Oil and Analysis of a Potential Nutritional Concentrate.

Ten liters of fatty acid ethyl esters were prepared according to example16 using safflower oil obtained at a local grocery. The esters were thenplaced in 2 L lots in a high vacuum vessel used to feed a 2″ wiped filmevaporator (Pope Scientific, Saukville Wis.). Vacuum (0.01 torr) wasapplied to the high vacuum flask to remove residual volatile materials.After vigorous bubbling had ceased the material was passed through thewiped film still at an initial high rate (20 mL/min) to removelow-boiling materials. The walls of the still were heated to 80 C forthis process. During evaporation vapors were condensed by traps chilledwith liquid nitrogen. After removing removing volatiles from the methylester solution the still was heated to 140 C and the ethyl esters werere introduced and the vacuum was maintained. The flow of liquid wasadjusted so that the flow of distillate was approximately 10 times theflow of residue. During this time 1 L of residue was collected. Theresidue of distillation was introduced to the still and afterdistillation under the same conditions a concentrate of 50 mL wasobtained.

Analysis of the ethyl esters and the safflower oil with high fieldproton NMR Spectroscopy (500 MHz Bruker, Milton, ON Canada) revealed twosmall but observable peaks in the spectrum contributed by plant sterolsand other triterpene alcohols. The distillate did not have observablesterol peaks. The residue of the distillation was also observed usinghigh-field proton NMR. The proton spectrum was comparable to a mixtureof ethyl esters with plant free and bound sterols and triterpenealcohols. With further preparation steps known to those skilled in theart the free and bound sterol fraction may be separated and used ascomponents of nutritional concentrates. The preparation may also be usedas a direct source of sterols.

Example 18 Recovery of Non Esterified Sterols From Canola Methyl EsterDistillate Residue

The residue of distillation obtained from Example 15 (0.50 g) was mixedwith KOH (0.3 g) dissolved in ethanol (2.5 mL) and water (2.5 mL) Themixture was heated at 65 C for 3 hours after which the ethanol wasremoved under vacuum. The resulting residue was diluted with water (15mL) and the unsaponifiable matter was extracted with petroleum ether(3×15 mL). The combined organic phases were dried over anhydrous sodiumsulphate. Evaporation of the petroleum ether under reduced pressure gavea white solid (158 mg). A portion of the solid was dissolved indeuterated chloroform and placed in an NMR tube. Analysis of the solidwith high field proton NMR Spectroscopy (500 MHz Bruker, Milton, ONCanada) revealed that the solid was primarily a mixture of the freealcohol forms of phytosterol compounds.

Example 19 Separation of Fractions From the Canola Methyl EsterDistillate Residue by Silica Chromatography

Fifteen grams of silica gel 60 was packed in a 10 cm glass column andthe column was washed with 50 mL of n-hexane. The wash was discarded.Canola methyl ester distillate residue (0.4 g) was dissolved in hexaneand added to the column. The column was then washed sequentially with 50mL of n-hexane, 50 mL of 3% diethyl ether in n-hexane, 50 mL of 10percent ethyl acetate in n-hexane and, finally, 50 mL of 25% ethylacetate in n-hexane. The repeated extractions produced four fractionswith masses of 250 mg, 50 mg, 10 mg and 65 mg respectively after thecomplete removal of the extraction solvent. The first three fractionswere oil like in nature while the last fraction was a white solid.Desolventized samples were dissolved in deuterated chloroform and placedin NMR tubes for analysis. Analysis of the fractions with high fieldproton NMR Spectroscopy (500 MHz Bruker, Milton, ON Canada) revealedthat the fractions were 1) a mixture of sterol esters of fatty acidswith some fatty acid methyl ester; 2) a mixture of fatty acid methylesters with some sterol ester; 3) a complex mixture containing Fattyacid esters as well as some unknown compounds; 4) A highly enrichedfraction of phytosterols in a free alcohol form.

1. A process for extracting lubricity enhancing and biologically activecompounds from oils, fats and greases, derived from animal and plantsources, comprising pressing a solid source material so as to releaseoils, fats and greases with lower levels of lubricity enhancingcompounds, and solvent extracting pressed solid source material so as toproduce a first oil, fat and grease concentrate of lubricity enhancingand biologically active compounds therefrom.
 2. A process according toclaim 1 including the steps of separating said lower level of lubricityenhancing and biologically active compounds from said pressed solidsource material and mechanically extracting a second oil, fat or greaseconcentrate with elevated levels of lubricity enhancing and biologicallyactive compounds.
 3. A process according to claim 2 including combiningsaid first and second concentrates.
 4. A process according to claim 1wherein triacyl glycerol molecules present in the concentrate arechemically modified so as to lower average molecular weight anddistillation is utilized to extract modified triglyceride products andleave a concentrate of lubricity enhancing and biologically activecompounds.
 5. A process according to claim 3 where triacyl glycerolmolecules present in the concentrates are chemically modified to lowerthe average molecular weight and distillation is utilized to extract themodified triglyceride products and leave a concentrate of lubricityenhancing compounds.
 6. A process according to claim 1 where the fatsare from tall oil.
 7. A process according to claim 1 wherein the plantsource is selected from the group consisting of soybean, canola, palm,olive, hemp, sunflower, rapeseed, flaxseed, corn and coconut.
 8. Aprocess according to claim 1 wherein the animal source is selected fromthe group consisting of swine, poultry and beef.
 9. A lubricityenhancing concentrate, derived by extraction from a source selected fromanimal and plant oils, fats and greases, and enriched in dolichol,dolichol phosphate, phospholipids, phospholipid metal ion complexes,diacylglycerides, monoacylglycerides, sterols, sterol fatty acyl esters,tocopherols, squalene, polyprenols, n-alkanols and wax esters.
 10. Aprocess according to claim 1 where triacyl glycerol molecules present inthe source material grease are converted to alkyl esters, alcohols,amides, alkanes, aldehydes, fatty acids or amines to lower the averagemolecular weight prior to distillation for preparation of the lubricityconcentrate.
 11. A process according to claim 3 where triacyl glycerolmolecules present in the source material are converted to alkyl esters,alcohols, amides, alkanes, aldehydes, fatty acids or amines to lower theaverage molecular weight prior to distillation for preparation of thelubricity concentrate.
 12. A process according to claim 1 where highmolecular weight substances are separated from lower molecular weightsubstances by size exclusion chromatography.
 13. A process according toclaim 1 where high molecular weight substances are separated from lowermolecular weight substances by crystallization.
 14. A process accordingto claim 1 where high molecular weight substances are separated fromlower molecular substances by any combination of distillation,crystallization and chromatography.
 15. A process for enhancinglubricity characteristics of kerosene comprising adding thereto alubricity enhancing concentrate as claimed in claim
 9. 16. A process forenhancing lubricity characteristics of diesel fuel comprising addingthereto a lubricity enhancing concentrate as claimed in claim
 9. 17. Aprocess for enhancing lubricity characteristics of jet fuel comprisingadding thereto a lubricity enhancing concentrate as claimed in claim 9.18. A process for enhancing lubricity characteristics of gasoline fuelfor internal combustion engines comprising adding thereto a lubricityenhancing concentrate as claimed in claim
 9. 19. A process for enhancinglubricity characteristics of motor oil comprising adding thereto alubricity enhancing concentrate as claimed in claim
 9. 20. A lubricityenhanced kerosene product comprising kerosene and a lubricity enhancingconcentrate as claimed in claim
 9. 21. A lubricity enhanced diesel fuelproduct comprising diesel fuel and a lubricity enhancing concentrate asclaimed in claim
 9. 22. A lubricity enhanced jet fuel product comprisingjet fuel and a lubricity enhancing concentrate as claimed in claim 9.23. A lubricity enhanced gasoline product comprising gasoline and alubricity enhancing concentrate as claimed in claim
 9. 24. A lubricityenhanced motor oil product comprising motor oil and a lubricityenhancing concentrate as claimed in claim
 9. 25. A biologically activeconcentrate derived by extraction from a source selected from animal andplant oils, fats and greases, and enriched in dolichol, dolicholphosphate, phospholipids, phospholipid metal ion complexes,diacylglycerides, monoacylglycerides, sterols, sterol fatty acyl esters,tocopherols, squalene, polyprenols, n-alkanols and wax esters.